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1.
Genes Immun ; 25(2): 158-167, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38570727

RESUMO

In this study, antibody response and a single-cell RNA-seq analysis were conducted on peripheral blood mononuclear cells from five different groups: naïve subjects vaccinated with AZD1222 (AZ) or Ad5-nCoV (Cso), individuals previously infected and later vaccinated (hybrid) with AZD1222 (AZ-hb) or Ad5-nCoV (Cso-hb), and those who were infected and had recovered from COVID-19 (Inf). The results showed that AZ induced more robust neutralizing antibody responses than Cso. The single-cell RNA data revealed a high frequency of memory B cells in the Cso and Cso-hb. In contrast, AZ and AZ-hb groups exhibited the highest proportion of activated naïve B cells expressing CXCR4. Transcriptomic analysis of CD4+ and CD8+ T cells demonstrated a heterogeneous response following vaccination, hybrid immunity, or natural infection. However, a single dose of Ad5-nCoV was sufficient to strongly activate CD4+ T cells (naïve and memory) expressing ANX1 and FOS, similar to the hybrid response observed with AZ. An interesting finding was the robust activation of a subset of CD8+ T cells expressing GZMB, GZMH, and IFNG genes in the Cso-hb group. Our findings suggest that both vaccines effectively stimulated the cellular immune response; however, the Ad5-nCoV induced a more robust CD8+ T-cell response in previously infected individuals.


Assuntos
Vacinas contra COVID-19 , COVID-19 , Humanos , Linfócitos T CD8-Positivos , Adenoviridae/genética , ChAdOx1 nCoV-19 , Leucócitos Mononucleares , Perfilação da Expressão Gênica , Imunidade Adaptativa , Anticorpos Neutralizantes/genética , Anticorpos Antivirais/genética
3.
iScience ; 26(4): 106562, 2023 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-37063467

RESUMO

This study reports the isolation and characterization of a human monoclonal antibody (mAb) called 19n01. This mAb was isolated by using single-cell RNAseq of B cells from donors infected with the ancestral strain. This mAb possesses a potent and broad capacity to bind and neutralize all previously circulating variants of concern (VOCs), including Omicron sublineages BA.1, BA.2, and BA.4/5. The pseudovirus neutralization assay revealed robust neutralization capacity against the G614 strain, BA.1, BA.2, and BA.4/5, with inhibitory concentration (IC50) values ranging from 0.0035 to 0.0164 µg/mL. The microneutralization assay using the G614 strain and VOCs demonstrated IC50 values of 0.013-0.267 µg/mL. Biophysical and structural analysis showed that 19n01 cross-competes with ACE2 binding to the receptor-binding domain (RBD) and the kinetic parameters confirmed the high affinity against the Omicron sublineages (KD of 61 and 30 nM for BA.2 and BA.4/5, respectively). These results suggest that the 19n01 is a remarkably potent and broadly reactive mAb.

4.
Front Immunol ; 14: 1276950, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38179057

RESUMO

Introduction: This study evaluated the immune response to a multiepitope recombinant chimeric protein (CHIVAX) containing B- and T-cell epitopes of the SARS-CoV-2 spike's receptor binding domain (RBD) in a translational porcine model for pre-clinical studies. Methods: We generated a multiepitope recombinant protein engineered to include six coding conserved epitopes from the RBD domain of the SARS-CoV-2 S protein. Pigs were divided into groups and immunized with different doses of the protein, with serum samples collected over time to determine antibody responses by indirect ELISA and antibody titration. Peptide recognition was also analyzed by Western blotting. A surrogate neutralization assay with recombinant ACE2 and RBDs was performed. Intranasal doses of the immunogen were also prepared and tested on Vietnamese minipigs. Results: When the immunogen was administered subcutaneously, it induced specific IgG antibodies in pigs, and higher doses correlated with higher antibody levels. Antibodies from immunized pigs recognized individual peptides in the multiepitope vaccine and inhibited RBD-ACE2 binding for five variants of concern (VOC). Comparative antigen delivery methods showed that both, subcutaneous and combined subcutaneous/intranasal approaches, induced specific IgG and IgA antibodies, with the subcutaneous approach having superior neutralizing activity. CHIVAX elicited systemic immunity, evidenced by specific IgG antibodies in the serum, and local mucosal immunity, indicated by IgA antibodies in saliva, nasal, and bronchoalveolar lavage secretions. Importantly, these antibodies demonstrated neutralizing activity against SARS-CoV-2 in vitro. Discussion: The elicited antibodies recognized individual epitopes on the chimeric protein and demonstrated the capacity to block RBD-ACE2 binding of the ancestral SARS-CoV-2 strain and four VOCs. The findings provide proof of concept for using multiepitope recombinant antigens and a combined immunization protocol to induce a neutralizing immune response against SARS-CoV-2 in the pig translational model for preclinical studies.


Assuntos
COVID-19 , Vacinas , Suínos , Animais , Humanos , Imunidade nas Mucosas , COVID-19/prevenção & controle , Vacinas contra COVID-19 , SARS-CoV-2 , Enzima de Conversão de Angiotensina 2 , Porco Miniatura , Epitopos de Linfócito T , Imunoglobulina A , Imunoglobulina G
5.
Nutrients ; 14(15)2022 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35893931

RESUMO

Food supplements of plant origin for weight control are increasingly being demanded by consumers as a way to promote good health. Among them, those based on Garcinia cambogia (GCFS) are widely commercialized considering their bioactive properties, mainly due to (-)-hydroxycitric acid ((-)-HCA). However, recently, controversy has arisen over their safety; thus, further research and continuous monitoring of their composition is required. Hence, in this work, a multi-analytical approach was followed to determine not only (-)-HCA but also other constituents of 18 GCFS, which could be used as quality markers to detect fraudulent practices in these samples. Discrepancies between the declared (-)-HCA content and that experimentally determined were detected by LC-UV in 33% of the samples. Moreover, GC-MS analyses of GCFS allowed the detection of different compounds not present in G. cambogia fruits and not declared on supplement labels, probably related to heat exposure or to the addition of excipients or other extracts. This multi-analytical methodology is shown to be advantageous to address different fraudulent practices affecting the quality of these supplements.


Assuntos
Fármacos Antiobesidade , Garcinia cambogia , Fármacos Antiobesidade/análise , Suplementos Nutricionais/análise , Frutas/química , Extratos Vegetais , Redução de Peso
6.
Appl Environ Microbiol ; 88(15): e0052622, 2022 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-35852362

RESUMO

The environmental context of the nitrogen-fixing mutualism between leguminous plants and rhizobial bacteria varies over space and time. Variation in resource availability, population density, and composition likely affect the ecology and evolution of rhizobia and their symbiotic interactions with hosts. We examined how host genotype, nitrogen addition, rhizobial density, and community complexity affected selection on 68 rhizobial strains in the Sinorhizobium meliloti-Medicago truncatula mutualism. As expected, host genotype had a substantial effect on the size, number, and strain composition of root nodules (the symbiotic organ). The understudied environmental variable of rhizobial density had a stronger effect on nodule strain frequency than the addition of low nitrogen levels. Higher inoculum density resulted in a nodule community that was less diverse and more beneficial but only in the context of the more selective host genotype. Higher density resulted in more diverse and less beneficial nodule communities with the less selective host. Density effects on strain composition deserve additional scrutiny as they can create feedback between ecological and evolutionary processes. Finally, we found that relative strain rankings were stable across increasing community complexity (2, 3, 8, or 68 strains). This unexpected result suggests that higher-order interactions between strains are rare in the context of nodule formation and development. Our work highlights the importance of examining mechanisms of density-dependent strain fitness and developing theoretical predictions that incorporate density dependence. Furthermore, our results have translational relevance for overcoming establishment barriers in bioinoculants and motivating breeding programs that maintain beneficial plant-microbe interactions across diverse agroecological contexts. IMPORTANCE Legume crops establish beneficial associations with rhizobial bacteria that perform biological nitrogen fixation, providing nitrogen to plants without the economic and greenhouse gas emission costs of chemical nitrogen inputs. Here, we examine the influence of three environmental factors that vary in agricultural fields on strain relative fitness in nodules. In addition to manipulating nitrogen, we also use two biotic variables that have rarely been examined: the rhizobial community's density and complexity. Taken together, our results suggest that (i) breeding legume varieties that select beneficial strains despite environmental variation is possible, (ii) changes in rhizobial population densities that occur routinely in agricultural fields could drive evolutionary changes in rhizobial populations, and (iii) the lack of higher-order interactions between strains will allow the high-throughput assessments of rhizobia winners and losers during plant interactions.


Assuntos
Medicago truncatula , Rhizobium , Genótipo , Medicago truncatula/genética , Medicago truncatula/microbiologia , Nitrogênio , Fixação de Nitrogênio/genética , Melhoramento Vegetal , Rhizobium/genética , Nódulos Radiculares de Plantas/microbiologia , Simbiose/genética
7.
Vaccines (Basel) ; 10(5)2022 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-35632440

RESUMO

Dendritic cell (DC) targeting by DEC205+ cells effectively promotes the internalization of antigens that may trigger a specific immune response. In this study, we evaluated the ability of a recombinant antibody, anti-DEC205 (rAb ZH9F7), to trigger cellular endocytosis in subpopulations of DCs and targeted cells after intradermal injection and subsequent migration toward lymph nodes. Furthermore, the cellular immune response was evaluated in pigs after intradermal application of the antigenized rAb ZH9F7 combined with porcine circovirus type 2 cap antigen (rAb ZH9F7-Cap). We demonstrated that rAb ZH9F7 recognized conventional type 1 and 2 DCs from the blood and skin and monocytes. It promoted receptor-mediated endocytosis and migration of cDCs and moDCs toward regional lymph nodes. Intradermal application of rAb ZH9F7-Cap induced a higher frequency of IFN-γ-secreting CD4+CD8+ T lymphocytes and antibodies against Cap protein than that in the control group. In conclusion, the rAb ZH9F7-Cap system promoted the target of skin cDC1 and cDC2, provoking migration to the regional lymph nodes and inducing a Th1 response, as evidenced by the proliferation of double-positive CD4+CD8+ T cells, which correlates with an enhanced ability to target the cDC1 subset both in vitro and in vivo.

8.
Transbound Emerg Dis ; 69(4): e734-e745, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34655457

RESUMO

The SARS-CoV-2 virus was detected for the first time in December 2019 in Wuhan, China. Currently, this virus has spread around the world, and new variants have emerged. This new pandemic virus provoked the rapid development of diagnostic tools, therapies and vaccines to control this new disease called COVID-19. Antibody detection by ELISA has been broadly used to recognize the number of persons infected with this virus or to evaluate the response of vaccinated individuals. As the pandemic spread, new questions arose, such as the prevalence of antibodies after natural infection and the response induced by the different vaccines. In Mexico, as in other countries, mRNA and viral-vectored vaccines have been widely used among the population. In this work, we developed an indirect ELISA test to evaluate S1 antibodies in convalescent and vaccinated individuals. By using this test, we showed that IgG antibodies against the S1 protein of SARS-CoV-2 were detected up to 42 weeks after the onset of the symptoms, in contrast to IgA and IgM, which decreased 14 weeks after the onset of symptoms. The evaluation of the antibody response in individuals vaccinated with Pfizer-BioNTech and CanSinoBio vaccines showed no differences 2 weeks after vaccination. However, after completing the two doses of Pfizer-BioNTech and the one dose of CanSinoBio, a significantly higher response of IgG antibodies was observed in persons vaccinated with Pfizer-BioNTech than in those vaccinated with CanSinoBio. In conclusion, these results confirm that after natural infection with SARS-CoV-2, it is possible to detect antibodies for up to 10 months. Additionally, our results showed that one dose of the CanSinoBio vaccine induces a lower response of IgG antibodies than that induced by the complete scheme of the Pfizer-BioNTech vaccine.


Assuntos
COVID-19 , Vacinas Virais , Animais , Anticorpos Antivirais , COVID-19/prevenção & controle , COVID-19/veterinária , Imunoglobulina A , Imunoglobulina G , Imunoglobulina M , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus
9.
Ophthalmic Epidemiol ; 29(5): 483-490, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-34530684

RESUMO

PURPOSE: During the COVID-19 pandemic, healthcare workers (HCWs) are at a considerable risk of being infected with SARS-CoV-2; among them, HCWs from ophthalmology departments are more prone to develop severe symptoms. In Mexico City, the prevalence of SARS-CoV-2 infection among HCWs is 30%. The present work aims to describe the seroprevalence among HCWs at an Ophthalmological Reference Centre in Mexico City. METHODS: A self-report questionnaire, RT-PCR test and detection of serum IgG/IgM antibodies against SARS-CoV-2 were performed among HCWs at the Institute of Ophthalmology "Conde de Valenciana". RESULTS: A total of 169 HCWs participated in the study. None of the participants declared severe symptoms, and only 15% showed three or more symptoms. The results showed that 32% of the participants were RT-PCR+ (54/169), and 20% (35/169) presented IgG antibodies against SARS-CoV-2. Thirteen percent of the RT-PCR+ subjects were IgG positive, and 7.6% of the RT-PCR- participants were IgG positive. The presence of three or more symptoms correlated with the presence of IgG antibodies, as well as Ct values of < 32 (p < 0,05). CONCLUSION: Most of the HCW cohort showed mild symptoms, and 69% of the RT-PCR+ participants did not show IgG antibodies against SARS-CoV-2. Seroprevalence was significantly associated with the presentation of COVID-19-associated symptoms.


Assuntos
COVID-19 , Oftalmologia , COVID-19/epidemiologia , Estudos Transversais , Atenção à Saúde , Pessoal de Saúde , Humanos , Imunoglobulina G , Imunoglobulina M , Pandemias , SARS-CoV-2 , Estudos Soroepidemiológicos
10.
Rev. crim ; 64(2): 63-75, 2022.
Artigo em Espanhol | LILACS | ID: biblio-1417900

RESUMO

El presente artículo expone los argumentos que edificó la Corte Constitucional a la hora de revisar la acción pública de inconstitucionalidad que se presentó en contra del parágrafo 1 del artículo 8 de la Ley 1843 de 2017. La importancia de identificar la estructura argumentativa de la Corte Constitucional, permite entender de qué manera los presupuestos rectores del derecho fundamental al debido proceso son vinculantes, al momento de impedir que la extensión de la solidaridad en el pago de una sanción pecuniaria derivada del uso de las cámaras de fotodetección, ponga en riesgo la aplicación del principio de responsabilidad individual, la imputación, la tipicidad y la debida defensa.


This article sets out the arguments built by the Constitutional Court when reviewing the public action of unconstitutionality filed against paragraph 1 of Article 8 of Law 1843 of 2017. The importance of identifying the argumentative structure of the Constitutional Court, allows understanding how the guiding assumptions of the fundamental right to due process are binding, when preventing the extension of solidarity in the payment of a pecuniary sanction derived from the use of photodetection cameras, jeopardizing the application of the principle of individual responsibility, imputation, typicality and due defense.


Este artigo apresenta os argumentos construídos pelo Tribunal Constitucional ao rever a ação pública de inconstitucionalidade apresentada contra o parágrafo 1 do artigo 8 da Lei 1843 de 2017. A importância de identificar a estrutura argumentativa do Tribunal Constitucional, nos permite compreender como são vinculativas as premissas norteadoras do direito fundamental ao devido processo, ao impedir a extensão da solidariedade no pagamento de uma penalidade financeira derivada do uso de câmeras fotodetetoras, colocando em risco a aplicação do princípio da responsabilidade individual, imputação, criminalidade e devida defesa.


Assuntos
Humanos , Direitos Humanos , Colômbia , Constituição e Estatutos , Sanção , Jurisprudência
11.
Foods ; 10(2)2021 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-33562045

RESUMO

Despite the nutritional properties of alfalfa, its production is mainly for animal feed and it is undervalued as a food source. In this study, the valorization of alfalfa as a potential source of bioactive carbohydrates [inositols, α-galactooligosaccharides (α-GOS)] is presented. A Box-Behnken experimental design was used to optimize the extraction of these carbohydrates from leaves, stems, and seeds of alfalfa by solid-liquid extraction (SLE) and microwave-assisted extraction (MAE). Optimal extraction temperatures were similar for both treatments (40 °C leaves, 80 °C seeds); however, SLE required longer times (32.5 and 60 min vs. 5 min). In general, under similar extraction conditions, MAE provided higher yields of inositols (up to twice) and α-GOS (up to 7 times); hence, MAE was selected for their extraction from 13 alfalfa samples. Pinitol was the most abundant inositol of leaves and stems (24.2-31.0 mg·g-1 and 15.5-22.5 mg·g-1, respectively) while seed extracts were rich in α-GOS, mainly in stachyose (48.8-84.7 mg·g-1). In addition, inositols and α-GOS concentrations of lyophilized MAE extracts were stable for up to 26 days at 50 °C. These findings demonstrate that alfalfa is a valuable source of bioactive carbohydrates and MAE a promising alternative technique to obtain functional extracts.

12.
Electrophoresis ; 41(20): 1804-1811, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32885861

RESUMO

A microwave-assisted extraction (MAE) method was developed for the extraction of bioactive inositols (D-chiro- and myo-inositols) from lettuce (Lactuca sativa) leaves as a strategy for the revalorization of these agrofood residues. Gas chromatography-mass spectrometry was selected for the simultaneous determination of inositols and sugars (glucose, fructose, and sucrose) in these samples. A Box-Behnken experimental design was used to maximize the extraction of inositols based on the results of single factor tests. Optimal conditions of the extraction process were as follows: liquid-to-solid ratio of 100:1 v/w, 40°C, 30 min extraction time, 20:80 ethanol:water (v/v), and one extraction cycle. When compared with conventional solid-liquid extraction (SLE), MAE was found to be more effective for the extraction of target bioactive carbohydrates (MAE 5.42 mg/g dry sample versus SLE 4.01 mg/g dry sample). Then, MAE methodology was applied to the extraction of inositols from L. sativa leaves of different varieties (var. longifolia, var. capitata and var. crispa). D-chiro- and myo-inositol contents varied between 0.57-7.15 and 0.83-3.48 mg/g dry sample, respectively. Interfering sugars were removed from the extracts using a biotechnological procedure based on the use of Saccharomyces cerevisiae for 24 h. The developed methodology was a good alternative to classical procedures to obtain extracts enriched in inositols from lettuce residues, which could be of interest for the agrofood industry.


Assuntos
Fracionamento Químico/métodos , Inositol/análise , Inositol/isolamento & purificação , Lactuca/química , Agricultura , Indústria Alimentícia , Cromatografia Gasosa-Espectrometria de Massas , Resíduos Industriais , Inositol/química , Micro-Ondas
13.
Plant Physiol ; 182(1): 463-471, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31653715

RESUMO

Genetic studies of legume symbiosis with nitrogen-fixing rhizobial bacteria have traditionally focused on nodule and nitrogen-fixation phenotypes when hosts are inoculated with a single rhizobial strain. These approaches overlook the potential effect of host genes on rhizobial fitness (i.e. how many rhizobia are released from host nodules) and strain-specific effects of host genes (i.e. genome × genome interactions). Using Medicago truncatula mutants in the recently described nodule-specific PLAT domain (NPD) gene family, we show how inoculating plants with a mixed inoculum of 68 rhizobial strains (Ensifer meliloti) via a select-and-resequence approach can be used to efficiently assay host mutants for strain-specific effects of late-acting host genes on interacting bacteria. The deletion of a single NPD gene (npd2) or all five members of the NPD gene family (npd1-5) differentially altered the frequency of rhizobial strains in nodules even though npd2 mutants had no visible nodule morphology or N-fixation phenotype. Also, npd1-5 nodules were less diverse and had larger populations of colony-forming rhizobia despite their smaller size. Lastly, NPD mutations disrupt a positive correlation between strain fitness and wild-type host biomass. These changes indicate that the effects of NPD proteins are strain dependent and that NPD family members are not redundant with regard to their effects on rhizobial strains. Association analyses of the rhizobial strains in the mixed inoculation indicate that rhizobial genes involved in chromosome segregation, cell division, GABA metabolism, efflux systems, and stress tolerance play an important role in the strain-specific effects of NPD genes.


Assuntos
Medicago truncatula/metabolismo , Nódulos Radiculares de Plantas/metabolismo , Medicago truncatula/genética , Medicago truncatula/microbiologia , Fixação de Nitrogênio/genética , Fixação de Nitrogênio/fisiologia , Nodulação/genética , Nodulação/fisiologia , Nódulos Radiculares de Plantas/genética , Nódulos Radiculares de Plantas/microbiologia , Sinorhizobium meliloti/fisiologia , Simbiose/genética , Simbiose/fisiologia
14.
New Phytol ; 222(3): 1538-1550, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30664233

RESUMO

Symbiotic nitrogen fixation in legumes is mediated by an interplay of signaling processes between plant hosts and rhizobial symbionts. In legumes, several secreted protein families have undergone expansions and play key roles in nodulation. Thus, identifying lineage-specific expansions (LSEs) of nodulation-associated genes can be a strategy to discover candidate gene families. Using bioinformatic tools, we identified 13 LSEs of nodulation-related secreted protein families, each unique to either Glycine, Arachis or Medicago lineages. In the Medicago lineage, nodule-specific Polycystin-1, Lipoxygenase, Alpha Toxin (PLAT) domain proteins (NPDs) expanded to five members. We examined NPD function using CRISPR/Cas9 multiplex genome editing to create Medicago truncatula NPD knockout lines, targeting one to five NPD genes. Mutant lines with differing combinations of NPD gene inactivations had progressively smaller nodules, earlier onset of nodule senescence, or ineffective nodules compared to the wild-type control. Double- and triple-knockout lines showed dissimilar nodulation phenotypes but coincided in upregulation of a DHHC-type zinc finger and an aspartyl protease gene, possible candidates for the observed disturbance of proper nodule function. By postulating that gene family expansions can be used to detect candidate genes, we identified a family of nodule-specific PLAT domain proteins and confirmed that they play a role in successful nodule formation.


Assuntos
Medicago truncatula/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Nodulação , Nódulos Radiculares de Plantas/metabolismo , Sequência de Aminoácidos , Regulação da Expressão Gênica de Plantas , Genótipo , Medicago truncatula/genética , Medicago truncatula/microbiologia , Fenótipo , Nodulação/genética , Domínios Proteicos , Rhizobium/fisiologia , Nódulos Radiculares de Plantas/microbiologia
15.
Enferm Clin (Engl Ed) ; 28(1): 36-43, 2018.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-29153438

RESUMO

OBJECTIVE: To determine the effect of the programme Hospitalisation discharge plan for patients with chronic diseases and family caregivers to strengthen their home care competence -CUIDAR- and reduce the caregiver burden. METHOD: Quasi-experimental study approach with intervention group. Participants were 62 patients and their caregivers who consulted with a health care institution in the city of Girardot (Colombia). The intervention was carried out for one month, during which, measurements were performed before and after the programme. RESULTS: Most patients were elderly, diagnosed with diabetes, hypertension or COPD, 35% of them with some degree of dependency. The caregivers were mostly women, between the ages of 35 and 59 years old, domestic caregivers, and responsible for caring for their patients for between 13-24hours a day. At the start of the programme the competency for care was low in both patients and caregivers, after the intervention there was a general increase in care and a statistically significant change. Also, at the beginning, 48% of caregivers had some level of burden, and after the study only 27% reported burden with care. CONCLUSIONS: The Hospitalisation discharge plan is a strategy that increases the home care competency of the patient and the caregiver, and decreases the caregiver burden.


Assuntos
Cuidadores , Doença Crônica , Assistência Domiciliar , Alta do Paciente , Adolescente , Adulto , Idoso , Efeitos Psicossociais da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
16.
Plant Physiol ; 173(2): 921-931, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28057894

RESUMO

Genome-wide association (GWA) studies offer the opportunity to identify genes that contribute to naturally occurring variation in quantitative traits. However, GWA relies exclusively on statistical association, so functional validation is necessary to make strong claims about gene function. We used a combination of gene-disruption platforms (Tnt1 retrotransposons, hairpin RNA-interference constructs, and CRISPR/Cas9 nucleases) together with randomized, well-replicated experiments to evaluate the function of genes that an earlier GWA study in Medicago truncatula had identified as candidates contributing to variation in the symbiosis between legumes and rhizobia. We evaluated ten candidate genes found in six clusters of strongly associated single nucleotide polymorphisms, selected on the basis of their strength of statistical association, proximity to annotated gene models, and root or nodule expression. We found statistically significant effects on nodule production for three candidate genes, each validated in two independent mutants. Annotated functions of these three genes suggest their contributions to quantitative variation in nodule production occur through processes not previously connected to nodulation, including phosphorous supply and salicylic acid-related defense response. These results demonstrate the utility of GWA combined with reverse mutagenesis technologies to discover and validate genes contributing to naturally occurring variation in quantitative traits. The results highlight the potential for GWA to complement forward genetics in identifying the genetic basis of ecologically and economically important traits.


Assuntos
Estudo de Associação Genômica Ampla , Medicago truncatula/genética , Nodulação/genética , Locos de Características Quantitativas/genética , Alelos , Sequência de Aminoácidos , Sequência de Bases , Mapeamento Cromossômico , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Genoma de Planta , Mutagênese/genética , Mutação/genética , Nitrogênio/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Reprodutibilidade dos Testes
17.
G3 (Bethesda) ; 4(10): 2003-12, 2014 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-25155275

RESUMO

The LEED..PEED (LP) gene family in Medicago truncatula (A17) is composed of 13 genes coding small putatively secreted peptides with one to two conserved domains of negatively charged residues. This family is not present in the genomes of Glycine max, Lotus japonicus, or the IRLC species Cicer arietinum. LP genes were also not detected in a Trifolium pratense draft genome or Pisum sativum nodule transcriptome, which were sequenced de novo in this study, suggesting that the LP gene family arose within the past 25 million years. M. truncatula accession HM056 has 13 LP genes with high similarity to those in A17, whereas M. truncatula ssp. tricycla (R108) and M. sativa have 11 and 10 LP gene copies, respectively. In M. truncatula A17, 12 LP genes are located on chromosome 7 within a 93-kb window, whereas one LP gene copy is located on chromosome 4. A phylogenetic analysis of the gene family is consistent with most gene duplications occurring prior to Medicago speciation events, mainly through local tandem duplications and one distant duplication across chromosomes. Synteny comparisons between R108 and A17 confirm that gene order is conserved between the two subspecies, although a further duplication occurred solely in A17. In M. truncatula A17, all 13 LPs are exclusively transcribed in nodules and absent from other plant tissues, including roots, leaves, flowers, seeds, seed shells, and pods. The recent expansion of LP genes in Medicago spp. and their timing and location of expression suggest a novel function in nodulation, possibly as an aftermath of the evolution of bacteroid terminal differentiation or potentially associated with rhizobial-host specificity.


Assuntos
Genoma de Planta , Medicago sativa/genética , Medicago truncatula/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Cromossomos de Plantas/genética , Cromossomos de Plantas/metabolismo , Medicago sativa/classificação , Medicago truncatula/classificação , Dados de Sequência Molecular , Pisum sativum/genética , Filogenia , Proteínas de Plantas/metabolismo , Alinhamento de Sequência , Transcriptoma
18.
Rev. salud pública ; 16(4): 600-611, jul.-ago. 2014. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-735161

RESUMO

Objetivo La situación de malnutrición afecta la mayor parte las zonas rurales de Colombia, involucrando principalmente la comunidad escolar. El objetivo de este estudio fue evaluar la situación nutricional de los estudiantes de la Institución Educativa Agrícola del municipio de Argelia (n=1528) ubicado en el suroccidente colombiano en el Departamento del Cauca. Métodos Se tomaron medidas de peso y talla y se determinó el estado nutricional a partir del índice de masa corporal (IMC) de acuerdo a los criterios definidos por la Organización Mundial de la Salud (OMS). Las mediciones antropométricas fueron analizadas a través de medidas de tendencia central estratificadas por género y rangos de edad. Para precisar la significancia estadística de algunas de las diferencias más importantes observadas entre los indicadores del estudio, se calculó el "error estándar de la diferencia" y se verificó tal significancia al nivel de 0.05. Resultados A partir del análisis del peso (P/E) y talla para la edad (T/E) e índice de masa corporal (IMC) se estableció una prevalencia de desnutrición y delgadez en niños y niñas entre 5 a 10 años, una tendencia hacia la obesidad en adolescentes y retraso en crecimiento tanto en niños como en adolescentes. Conclusiones Los resultados sugieren promoción de actividades deportivas a temprana edad, capacitación en hábitos de alimentación saludable a los escolares y sus familias, y se constituyen en un soporte científico para la planeación de estrategias de prevención y control de las entidades estatales de salud.(AU)


Objective Malnutrition affects most rural areas in Colombian, mainly involving the school-aged population. The aim of this study was to assess the nutritional status of students (n=1,528) attending an Agricultural College in the town of Argelia in south-western Colombia in the Cauca department. Methods The school-aged children’s weight and height were measured and their nutritional status was determined from their body mass index (BMI), according to criteria defined by the World Health Organization (WHO). Anthropometric measurements were analyzed by means of measures of central tendency, stratified by gender and age. The standard error of the difference was estimated to specify the statistical significance of some of the most important differences observed between study indicators; such significance was verified at 0.05 level. Results Prevalence regarding malnutrition and thinness in 5-10 years-old children, a trend towards obesity in adolescents and growth retardation in children and adolescents was established from analysis of the children’s weight (P/E), height for age (H/A) and BMI. Conclusions The results suggested that sports should be encouraged at an early age, as well as training school children and their families regarding healthy eating habits and thus provide scientific support for planning government healthcare agencies’ prevention and control strategies.(AU)


Assuntos
Humanos , Pré-Escolar , Criança , Estado Nutricional/fisiologia , Desnutrição/epidemiologia , Índice de Massa Corporal , Avaliação Nutricional , Colômbia/epidemiologia
19.
Rev Salud Publica (Bogota) ; 16(4): 547-59, 2014.
Artigo em Espanhol | MEDLINE | ID: mdl-25791306

RESUMO

OBJECTIVE: Malnutrition affects most rural areas in Colombian, mainly involving the school-aged population. The aim of this study was to assess the nutritional status of students (n=1,528) attending an Agricultural College in the town of Argelia in south-western Colombia in the Cauca department. METHODS: The school-aged children’s weight and height were measured and their nutritional status was determined from their body mass index (BMI), according to criteria defined by the World Health Organization (WHO). Anthropometric measurements were analyzed by means of measures of central tendency, stratified by gender and age. The standard error of the difference was estimated to specify the statistical significance of some of the most important differences observed between study indicators; such significance was verified at 0.05 level. RESULTS: Prevalence regarding malnutrition and thinness in 5-10 years-old children, a trend towards obesity in adolescents and growth retardation in children and adolescents was established from analysis of the children’'s weight (P/E), height for age (H/A) and BMI. CONCLUSIONS: The results suggested that sports should be encouraged at an early age, as well as training school children and their families regarding healthy eating habits and thus provide scientific support for planning government healthcare agencies’ prevention and control strategies.


Assuntos
Transtornos do Crescimento/epidemiologia , Desnutrição/epidemiologia , Estado Nutricional , Obesidade/epidemiologia , Saúde da População Rural/estatística & dados numéricos , Magreza/epidemiologia , Adolescente , Índice de Massa Corporal , Criança , Pré-Escolar , Colômbia/epidemiologia , Estudos Transversais , Feminino , Transtornos do Crescimento/diagnóstico , Humanos , Masculino , Desnutrição/diagnóstico , Avaliação Nutricional , Obesidade/diagnóstico , Prevalência , Magreza/diagnóstico , Adulto Jovem
20.
Water Res ; 47(10): 3282-90, 2013 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-23602619

RESUMO

The haloacetic acids (HAAs) are the second-most prevalent class of drinking water disinfection by-products formed by chemical disinfectants. Previous studies have determined DNA damage and repair of HAA-induced lesions in mammalian and human cell lines; however, little is known of the genomic DNA and chromosome damage induced by these compounds in primary human cells. The aim of this study was to evaluate the genotoxic and clastogenic effects of the monoHAA disinfection by-products in primary human lymphocytes. All monoHAAs were genotoxic in primary human lymphocytes, the rank order of genotoxicity and cytotoxicity was IAA > BAA >> CAA. After 6 h of repair time, only 50% of the DNA damage (maximum decrease in DNA damage) was repaired compared to the control. This demonstrates that primary human lymphocytes are less efficient in repairing the induced damage by monoHAAs than previous studies with mammalian cell lines. In addition, the monoHAAs induced an increase in the chromosome aberration frequency as a measurement of the clastogenic effect of these compounds. These results coupled with genomic technologies in primary human cells and other mammalian non-cancerous cell lines may lead to the identification of biomarkers that may be employed in feedback loops to aid water chemists and engineers in the overall goal of producing safer drinking water.


Assuntos
Desinfetantes/toxicidade , Desinfecção/métodos , Água Potável/química , Linfócitos/efeitos dos fármacos , Mutagênicos/toxicidade , Acetatos/química , Acetatos/toxicidade , Adulto , Células Cultivadas , Aberrações Cromossômicas , Dano ao DNA/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Desinfetantes/química , Humanos , Ácido Iodoacético/química , Ácido Iodoacético/toxicidade , Masculino , Índice Mitótico , Testes de Mutagenicidade
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